Fig. 1

Microglia morphology changes upon activation with cytokines. (A) Representative images of C20 microglia in -CTL group (unstimulated) and +CTL group (stimulated with 5ng/mL each of IFN-γ and TNF-α). Actin, Golgi & plasma membrane (PM) (Red, Phalloidin/WGA), Nuclei (Blue, Hoechst): Scale bar=100μm (B) Individual cellular features (i) Perimeter, (ii) Form Factor and (iii) Aspect Ratio and (iv) Principal component 1 (PC1_morpho) calculated using 21 features. Each point is a median of ~700-1000 cells/well with mean and standard deviation plotted for n=8 wells per group. *p<0.01 vs -CTL for all groups conducted using unpaired t-test with Welch’s correction. (C, D) Experimental and Operator variability: Microglia controls (-CTL vs +CTL) are significantly different across multiple experiments and operators. p<0.001 vs +CTL for all experiments using 2-way ANOVA with Sidak’s multiple comparisons test. (E) Z-factor for PC1_morpho is > 0.5 (dotted line) for 7/8 experiments